Targeting and glycosylation of patatin the major potato tuber protein in leaves of transgenic tobacco |
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Authors: | Uwe Sonnewald Arnd Sturm Maarten J Chrispeels Lothar Willmitzer |
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Institution: | (1) Institut für Genbiologische Forschung Berlin GmbH, Ihnestrasse 63, D-1000 Berlin 33;(2) Department of Biology, C-016 University of California, 92093 San Diego, La Jolla, CA, USA;(3) Present address: Friedrich Miescher Institute, P.O. Box 2543, CH-4002 Basel, Switzerland |
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Abstract: | Patatin, the most abundant protein in the storage parenchyma cells of potato (Solanum tuberosum L.) tubers, is a vacuolar glycoprotein that consists of a number of closely related polypeptides and is encoded by a large gene family. To analyse the glycosylation pattern and the nature of the glycans on a single patatin polypeptide in a heterologous tissue we introduced a single chimaeric patatin gene into tobacco (Nicotiana tabacum L.) and studied its product in leaves. Patatin isolated from the leaves of transgenic tobacco plants is glycosylated at asparagine (Asn)60, and Asn90, but the third glycosylation site (Asn202) has no glycan. The two glycans are typical small complex glycans with xylose, fucose, mannose and N-acetylglucosamine in a ratio 1:1:3:2, the same ratio as found on patatin isolated from potato tubers. Expression of patatin in tobacco leaves was accompanied by the correct processing of the signal peptide, and the proper targeting of the glyco-protein to the vacuoles of mesophyll cells.Abbreviations Asn
asparagine
- ConA
concanavalin A
- EndoH
endoglycosidase H
- Fuc
fucose
- GlcNAc
N-acetylglucosamine
- HPLC
high-performance liquid chromatography
- Man
mannose
- PAGE
polyacrylamide gel electrophoresis
- SDS
sodium dodecyl-sulfate
- Ser
serine
- TFMS
trifluoromethanesulfonic acid
- Thr
threonine
- Xyl
xylose |
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Keywords: | Glycoprotein Nicotiana (transgenic) Patatin Protein targeting Tuber (protein) Vacuole |
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